The purpose of the present pilot study was to evaluate the effect of a hydrogel composedof hyaluronic acid (HA) and platelet-rich plasma (PRP) as a carrier for human mesenchymal stemcells (hMSCs) for intervertebral disc (IVD) regeneration using a disc organ culture model. HA wasmixed with batroxobin (BTX) and PRP to form a hydrogel encapsulating 1 × 106 or 2 × 106 hMSCs.Bovine IVDs were nucleotomized and filled with hMSCs suspended in ~200 µL of the PRP/HA/BTXhydrogel. IVDs collected at day 0 and nucleotomized IVDs with no hMSCs and/or hydrogel alonewere used as controls. hMSCs encapsulated in the hydrogel were also cultured in well plates toevaluate the effect of the IVD environment on hMSCs. After 1 week, tissue structure, scaffoldintegration, hMSC viability and gene expression of matrix and nucleus pulposus (NP) cell markerswere assessed. Histological analysis showed a better preservation of the viability of the IVD tissueadjacent to the gel in the presence of hMSCs (~70%) compared to the hydrogel without hMSCs.Furthermore, disc morphology was maintained, and the hydrogel showed signs of integration withthe surrounding tissues. At the gene expression level, the hydrogel loaded with hMSCs preservedthe normal metabolism of the tissue. The IVD environment promoted hMSC differentiation towardsa NP cell phenotype by increasing cytokeratin-19 (KRT19) gene expression. This study demonstratedthat the hydrogel composed of HA/PRP/BTX represents a valid carrier for hMSCs being able tomaintain a good cell viability while stimulating cell activity and NP marker expression.

A hyaluronan and platelet-rich plasma hydrogel for mesenchymal stem cell delivery in the intervertebral disc: An organ culture study

Russo F.;Papalia R.;Vadalà G.;Denaro V.
2021-01-01

Abstract

The purpose of the present pilot study was to evaluate the effect of a hydrogel composedof hyaluronic acid (HA) and platelet-rich plasma (PRP) as a carrier for human mesenchymal stemcells (hMSCs) for intervertebral disc (IVD) regeneration using a disc organ culture model. HA wasmixed with batroxobin (BTX) and PRP to form a hydrogel encapsulating 1 × 106 or 2 × 106 hMSCs.Bovine IVDs were nucleotomized and filled with hMSCs suspended in ~200 µL of the PRP/HA/BTXhydrogel. IVDs collected at day 0 and nucleotomized IVDs with no hMSCs and/or hydrogel alonewere used as controls. hMSCs encapsulated in the hydrogel were also cultured in well plates toevaluate the effect of the IVD environment on hMSCs. After 1 week, tissue structure, scaffoldintegration, hMSC viability and gene expression of matrix and nucleus pulposus (NP) cell markerswere assessed. Histological analysis showed a better preservation of the viability of the IVD tissueadjacent to the gel in the presence of hMSCs (~70%) compared to the hydrogel without hMSCs.Furthermore, disc morphology was maintained, and the hydrogel showed signs of integration withthe surrounding tissues. At the gene expression level, the hydrogel loaded with hMSCs preservedthe normal metabolism of the tissue. The IVD environment promoted hMSC differentiation towardsa NP cell phenotype by increasing cytokeratin-19 (KRT19) gene expression. This study demonstratedthat the hydrogel composed of HA/PRP/BTX represents a valid carrier for hMSCs being able tomaintain a good cell viability while stimulating cell activity and NP marker expression.
2021
Ex vivo
Hyaluronic acid
Intervertebral disc
Mesenchymal stem cells
Organ culture
Platelet-rich plasma
Animals
Batroxobin
Cattle
Cell Differentiation
Gene Expression Regulation
Humans
Hyaluronic Acid
Hydrogels
Intervertebral Disc
Intervertebral Disc Degeneration
Keratin-19
Mesenchymal Stem Cells
Nucleus Pulposus
Organ Culture Techniques
Platelet-Rich Plasma
Mesenchymal Stem Cell Transplantation
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/20.500.12610/65562
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